Team:Washington/Protocols/PCR

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Revision as of 21:43, 13 September 2011

    1. Setup Amplification PCR Reaction
      1. 1uL Template
      2. 1uL 25mM dNTP's
      3. 10uL Phusion HF Buffer
      4. 0.5uL Forward Primer (Tm 65)
      5. 0.5uL Reverse Primer (Tm 65)
      6. 0.5uL Phusion polymerase
      7. 36.5uL diH2O
    2. Amplification PCR Reaction
      1. 98C - 30s
      2. 98C - 10s
      3. 63C - 10s
      4. 72C - 30s/kb target gene
      5. Repeat 2-4 29x
      6. 72C - 5min
      7. 10C - forever
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